DE102014106916A1 - Device for the automated determination of at least two different process parameters - Google Patents

Abstract

The invention relates to a device for the automated determination of at least two different process parameters of a process liquid of a process, in particular a bioprocess, comprising: a first measuring cell which is designed to provide a first measurement signal dependent on a first process parameter of a first sample of the process liquid; A second measuring cell which is designed to provide a second measuring signal which is dependent on a second process parameter of a second sample of the process liquid; and a control and evaluation device which serves to monitor and / or control the process and which is designed to receive and process the first and the second measurement signal, in particular to determine a measurement value of the first process parameter on the basis of the first measurement signal and determine a measured value of the second process parameter on the basis of the second measurement signal; wherein the first measurement signal and the second measurement signal serve different functions in the context of monitoring and / or controlling the process. The first process parameter can be a critical process parameter (CPP), and the second process parameter can be a critical quality attribute (CQA) parameter of the process.

Description

The invention relates to a device and a method for the automated determination of at least two different process parameters in a sample of a process fluid of a process. The process can be a bioprocess. In particular, the invention relates to an apparatus and a method for the automated determination of a control parameter, e.g. a metabolite content, and a product quality relevant parameter, e.g. a target protein content, of samples of the process fluid.

The ultimate goal of (bio) process analytics, particularly within the US Food and Drug Administration (PAT) Guidelines, is to improve productivity while maintaining consistent quality characteristics under the pressure of reducing time-to-market. The abbreviation PAT stands for "Process Analytical Technology". The PAT Guide was created by the FDA as an incentive and tool to optimize, analyze and control pharmaceutical manufacturing processes. According to this guideline, the critical process parameters influencing the critical quality characteristics of a biopharmaceutical manufacturing process must be analyzed and controlled. The critical process parameters (referred to in the PAT Guide as critical process parameters, CPP for short) are process variables of the process that enter into appropriate control algorithms for process control and regulation. In the following, these parameters are also referred to as control parameters. A distinction must be made between the parameters relevant to product quality (referred to in the PAT Guide as critical quality attributes, CQA for short). These parameters serve as a measure of product quality, but are not currently used as control parameters for the production process. The control parameters influence the product quality relevant parameters.

An example of the importance of bioprocess control is the production of recombinant proteins. Heterologous gene expression is induced only after reaching a specific cell density. Thus, within the biphasic culture process, both cell growth and the change to the metabolic phase in which product formation occurs must be closely monitored and controlled. Between cell growth and product production, which can not be directly correlated with one another depending on the process, a robust, reproducible process procedure / regime must be determined by optimizing the cultivation conditions. Such as in Rodrigues, ME, Costa, AR, Henriques, M. Azeredo, J., Oliveira, R. (2010). Technological progress in monoclonal antibody production systems. Biotechnol Prog, 26 (2), p.332-351 , One approach is to optimize the nutrient delivery strategy. For this purpose, nutrients, especially metabolic products (Metabolededukte) especially glucose, glutamine, and metabolites (metabolism products) such as Lactate, ammonium, glutamate, in general the metabolites, can be reliably determined (Rodrigues et al., P. , The term metabolite is understood here and below to mean not only a product or intermediate product of the metabolism, but more generally a substance involved in the metabolism, in particular metabolic products, intermediates and products. Too low but too high a metabolite concentration (increased formation of toxic products) can lead to a decrease in cell growth and / or productivity. Direct monitoring of the formed product would be extremely valuable and important for cell line selection and optimization of culture parameters ( Rodrigues et al. P. 343 right 4th paragraph ). A system which can precisely determine several critical process parameters within the same process-connected automation platform, one from the group of metabolites and the other from the group of specific products, is thus far more profitable.

The underlying concept of the PAT Guide is to control and / or regulate the process by defining appropriate control parameters that are collected online, thereby achieving a desired product quality in a more efficient manner. Essential for the control and regulation of bioprocesses within the meaning of the PAT Guide is therefore the existence or the development of suitable on-line sensor technology; i.e. Sensors and process-related analytical measuring technology.

Classical control parameters, in particular also for the control and / or regulation of bioprocesses, usually represent chemical / physical state variables such as temperature, pH, CO ₂ , O ₂ content, the determination of which is established or at least possible by means of inline sensor technology. With biosensor-based measuring systems, further control parameters can be determined that would not be accessible with classical, established measurement technology. Due to the instability of biological components, such systems have not proven to be suitable for inline routine use, so that for biosensor-based measuring systems it is necessary to take a sample and supply it to the measuring cell. Biosensor-based certain control parameters are, for example, metabolites, ie nutrients or metabolic products, whose content during a biological process has a direct influence on the process control in order to set the optimum conditions for product production - with the required quality.

The determination of the product quality-relevant parameters, the CQAs, provides a direct statement as to whether the product has the required properties within a defined tolerance range defined by the regulatory authorities, especially in pharmaceutical production. At present, samples taken manually from the process are time-displaced in the laboratory to determine the CQAs daily and a variety of product quality parameters are determined with appropriate laboratory analyzers. In most cases, all samples are examined together only after the process has ended. The examination of the samples requires trained personnel to operate and interpret the results. A reaction / action in case of non-compliance with the desired product properties is not possible with such a procedure.

Some methods for determining individual CQAs and CPPs as well as analysis devices of commercial providers for carrying out these methods are already known from the prior art. These are listed in the following Table I. In order for such devices to be used in automated process monitoring for monitoring or for controlling and / or regulating production processes, they require a process connection via which they can be connected to the process in order to carry out measurements or to remove samples from the process where the measurements can be made. In Table I is therefore indicated in each case whether the said analyzers have a process connection. Table I: method device Process connection CQA: product quantity CQA: product quality CPP: metabolite HPLC eg UltiMate 3000 HPLC Systems (Thermo Scientific Dionex, Thermo Fisher Scientific Inc., Waltham, MA, US) No Yes Yes No ELISA Mostly manually No Yes Yes No Amperometric, enzyme sensors YSI Flownamics SEG-FlOW (YSI Inc./Xylem Inc., Yellow Springs, Ohio, US) yes (for laboratory reactor) No No Yes Bioprofile Analyzer (Nova Biomedical Corporation, Waltham, MA, US) No No No Yes Biosensor Arrays (Jobst Technologies GmbH, Freiburg, DE) No No No Yes BioPAT Trace (Sartorius Stedim Biotech GmbH, Göttingen, DE) yes (laboratory) No No Yes photometry Konelab (Thermo Fisher Scientific, Inc., Waltham, MA, US), CuBiAn and Cedex Bio (F. Hoffmann-La Roche AG, Basle, CH) No yes (only immune gobuline) No Yes Separate laboratory equipment united under one control Baychromat Process / Lab (Bayer Technology Services GmbH, Leverkusen, DE) yes (laboratory and process) yes (only for Immununglobu line as product) No Yes

Out EP 1698891 A1 is a method for reducing the measurement deviation of amperometric biosensors known. This document refers to the modification of potential operation when an electrical mediator is used as a delivery mediator of the actual redox reaction of the analyte. The systematic error of the so-called background current, especially at the beginning of the reaction after storage before use, should be reduced by the claimed procedure.

In DE 3406223 A1 and US 3655958 an analyzer for automatically performing the standard addition method is described, the US patent reference to spectrophotometer takes.

Out US 2007/0224702 A1 describes a method for determining multiple analytes in one or more samples, each analyte being detected by a separate affinity assay.

In DE 196 12 766 A1 describes a method for the analysis of a complex biological system in which metabolic parameters are determined on the basis of samples taken from a fermenter by means of ligand-receptor interaction. The control of the process via sensors arranged in the fermenter.

Out DE 10 2010 064 391 A1 a method and an analyzer for the automated determination of an analyte content of a liquid sample is known, which basically also allows the detection of several different analytes in successive measurement cell supplied samples of a process liquid.

In US 2005/0208473 A1 A method for controlling a bioprocess is described in which various control parameters are detected by sensors arranged in the bioreactor, inter alia enzyme electrodes.

Out US 2007/0292958 A1 a device is known in which a process fluid bioprocess via a microdialysis probe (micro dialysis probe) are removed from a fermentation process and an analysis device for determining metabolite concentrations can be supplied.

In US 2008/0241966 A1 For example, a method and apparatus suitable for the automated determination of various metabolites in a fluid sample are described.

It is the object of the invention to provide a generic device or a method which avoids the disadvantages of the known from the prior art analyzers and methods. In particular, the device and the method should make it possible to monitor, control and / or regulate processes, in particular bioprocesses according to the PAT guide, and to save as much energy and resources as possible.

This object is achieved by the device according to claim 1 and the method according to claim 12. Advantageous embodiments are given in the dependent claims.

• – eine erste Messzelle, welche dazu ausgestaltet ist, ein von einem ersten Prozessparameter einer ersten Probe des Prozessmediums abhängiges erstes Messsignal bereitzustellen;
• – eine zweite Messzelle, welche dazu ausgestaltet ist, ein von einem zweiten Prozessparameter einer zweiten Probe des Prozessmediums abhängiges zweites Messsignal bereitzustellen; und
• – eine Steuerungs- und Auswertungseinrichtung, welche der Überwachung und/oder Steuerung des Prozesses dient, und welche dazu ausgestaltet ist, das erste und das zweite Messsignal zu empfangen und zu verarbeiten, insbesondere anhand des ersten Messsignals einen Messwert des ersten Prozessparameters zu bestimmen und anhand des zweiten Messsignals einen Messwert des zweiten Prozessparameters zu bestimmen; wobei das erste Messsignal und das zweite Messsignal unterschiedlichen Funktionen im Rahmen der Überwachung und/oder Steuerung des Prozesses dienen.

The device according to the invention for the automated determination of at least two different process parameters of a process medium of a process, in particular of a bioprocess, comprises:

• A first measuring cell, which is designed to provide a first measuring signal dependent on a first process parameter of a first sample of the process medium;
• A second measuring cell, which is designed to provide a second measuring signal dependent on a second process parameter of a second sample of the process medium; and
• A control and evaluation device which serves to monitor and / or control the process, and which is configured to receive and process the first and the second measurement signal, in particular to determine a measured value of the first process parameter based on the first measurement signal and based on the second measurement signal to determine a measured value of the second process parameter; wherein the first measurement signal and the second measurement signal serve different functions in the context of monitoring and / or controlling the process.

A measuring cell comprises a receiving space for receiving the respective sample, for example a container or a line, and at least one measuring sensor, which is designed to detect a measured variable of the measuring pickup of the measuring pickup of the sample recorded in the receiving space. The sensor may comprise a transducer, which is designed to convert the measured variable into an electrical signal which, optionally further processed, is output as a measuring signal. For example, the sensor may contact the sample by contacting the sample. alternative it may be in contact with the sample by detecting radiation emitted by the sample or detecting radiation incident on the sample after interacting with the sample.

The process medium may be, for example, a process fluid. The first and second samples may be formed from a predetermined amount of process liquid taken from the process container by dividing the withdrawn amount of liquid. It is also possible to remove the first and the second sample from the process container in succession.

By configuring the device to determine process-specific measuring signals, which serve different functions in the context of monitoring and / or controlling the process, and to make them available to the control and evaluation device, it is no longer necessary to use different measuring devices for determining different parameters of different functions often the measuring instruments also have an influence on parameters that are not relevant for the respective specific process. The measuring cells can be designed relatively simply and, in addition to the receiving space and the sensor, optionally only a suburb electronics for a first processing of the measurement signals, in particular for amplification and / or digitization include. In contrast to a measurement setup, in which different, complete measuring devices are combined with their own control and evaluation device with input means and display device, which each record a parameter, so a much more compact measurement setup is possible.

The use of a, in particular single, central control and evaluation device, which processes both measuring signals and controls both measuring cells, in particular both measuring sensors, permits a timely determination of the parameters and a comfortable operation of the device. In addition, the device is able to coordinate the determination of the parameters and their use for control tasks. This allows more energy and resource efficient monitoring, control and regulation of industrial processes and laboratory processes. If the higher-level control and evaluation device has input means and a display device, e.g. a display for displaying measured values, device parameters and possibly evaluation information, a simple and convenient operation of the device is possible, while in a measurement setup in which only a plurality of individually operated, complete, in particular via their own control and evaluation device with input - And output means equipped, measuring instruments are combined, the operation and the maintenance of the measurement setup are significantly more complex and trained personnel is required.

An embodiment is advantageous in which the first process parameter is a critical parameter (CPP), and the second process parameter is a product quality-relevant parameter (critical quality attribute, CQA) of the process.

While in the current state of the art, in which the CQAs with respect to the sampling time, especially in most cases only after the completion of the process, determined, it is advantageous to a selected or a few selected product quality parameters with a fully automated, process-based analysis system to determine a very small time offset for direct bioprocess monitoring. This does not necessarily mean that final controls of the product obtained during manufacture are not required in the laboratory to prove the specifications (final controls). In the case of timely determination of the product quality-relevant parameters, as provided here, it is advantageously possible to check compliance with the product specifications in parallel to the process management and, if the process is not complied with, to be stopped prematurely so that no further resources (in particular time and costs of the culture media) are consumed without product yield.

In the following Table II, some important product quality (CQA) and control parameters (CPP) to be determined in practice and possible biosensor-based measuring methods with optical and amperometric detection are listed by way of example, which can be determined with a flow measurement system. Table II:

To determine the product quality and control parameters with the device according to the invention, different measurement methods and detection principles can be used and combined with each other. Depending on the required parameters, established laboratory methods can be transferred for the automated sequence in the device according to the invention and / or commercially available reagent kits can be used. For the simple, robust, process-capable automation, a measurement procedure within a flow-through system is particularly advantageous. Within such a heterogeneous as well as homogeneous biological assays can be carried out and the measured variable suitable, mostly optically detected. Preferred here is a solid phase-bound affinity immunosensor with the in DE 201010064391 A1 . DE 102010064392 A1 . WO 2012055606 A1 and WO 2012055607 A1 described properties.

For example, analyte or other target molecules of the assay bound to immobilized receptors may be provided with an enzyme label so that when measured in a flow-through system, a substrate reacted with the enzyme to produce a product with altered optical properties (eg, absorption or fluorescence) is transported to the receptor-bound, enzyme-labeled analyte or target molecules and the product formed is transported away. Actual values of the product quality or control parameter to be determined may be obtained by evaluating the history of the primary signal of the optical measurement, e.g. an absorption measurement, be used in passing the substrate solution. Advantageously, the optical measurement is spatially separated, in particular downstream with respect to the detection area comprising the immobilized receptors.

In particular, metabolite contents or the proportion of viable or dead cells are suitable as control parameters. Product quality relevant parameters are activities, affinity properties or protein aggregation of the specific product. In a simple view, the content of the specific product may also be considered as CQA. In an activity determination as CQA, the proportion of the active substance (= biologically active substance) is determined. Particularly advantageous is the determination of the proportion of the active substance within the total content of the substance. This is e.g. coagulation factor 8 is possible on the basis of an activity determined by a homogeneous method (in particular in solution) together with a coagulation factor 8 content determined by means of a solid-phase affinity immunoassay.

The first process parameter mentioned above may be selected from the group consisting of: a metabolite content of the sample, in particular an anabolite or catabolite content, and a content of viable and / or dead cells.

The metabolite content may be a nutrient content or a metabolic product content. In particular, the metabolite content may include a glucose content, a lactate content, an ammonium content, a glutamine content, a lactose content, a galactose content, a sucrose content, a maltose content, an alanine content, an aspartic acid content, an I-amino acid content, an acetylcholine content, a pyruvate content, an acetate content, a citrate content.

The second process parameter may be selected from the group consisting of: the content of the product of the process in the liquid sample, an enzyme activity, a coagulation factor 8 content and activity, a content of specific immunoglobulin (Ig) types, in particular an IgG, IgA, IgM content, host cell protein content, the glycosylation pattern / structure of the product, the affinity of the analyte for a target, and protein aggregation.

In an advantageous embodiment of the invention, the first and / or the second measuring cell is configured to the corresponding measuring signal biosensor-based, i. by detecting the specific affinity recognition of the analyte by biological or biomolecular recognition structures. Advantageously, the first or the second measuring cell is designed to carry out a solid-phase, immunological assay for generating its measuring signal. In this embodiment, the respective other measuring cell can generate its measuring signal without carrying out a solid phase-bound immunological assay, in particular using an optical or an amperometric sensor and / or a photometric cuvette test. As already mentioned, both measuring cells can be operated in flow mode.

Advantageously, the first measuring cell for determining CPPs has at least one enzyme sensor.

The device may have a process connection via which it can be connected to a bioreactor. The process connection can in particular comprise an apparatus for automatic sampling from a process container in which the process liquid is contained and / or a device for sample treatment. The sampling device may be connectable to both laboratory bioreactors and process bioreactors. It can, for example, have a sample lock for connecting a liquid feed line to the bioreactor and a device for transporting process medium through the liquid feed line from the bioreactor in the direction of the measuring cells.

The device can advantageously be designed to carry out a standard addition method for determining the first or second parameter. For this purpose, a corresponding control algorithm can be stored in the control and evaluation device. In this embodiment, the device additionally comprises at least one container serving as a liquid reservoir with standard liquid, which can be supplied to the liquid sample to be supplied in the receptacle of the measuring cell or already contained in the receptacle of the measuring cell in order to displace the liquid sample with one or more predetermined volumes of the standard liquid , The addition of standard liquid to the liquid sample can be controlled by means of one or more pumps and / or valves controlled by the control and evaluation device.

The invention also relates to a method for the automated determination of at least two substances in samples of a process fluid by means of a device according to the embodiments described above, wherein the determination of the first and the second parameters are independent of each other.

The first parameter in this method may be a control parameter CPP, in particular a metabolite content, by means of a, in particular enzyme sensor, and as a second parameter a product quality-relevant parameter, e.g. a product level or product activity can be determined by means of a solid-phase, affinity assay.

The first and the second parameters can be determined several times (without the use of manual steps) during a bioprocessing process by repeatedly executable step sequences which involve the passage of one or more liquids through the measuring cells. In particular, the measurement signal of the first and / or second measuring cell can be generated in the flow-through method.

Thus, it is particularly in the case that the second measuring cell has enzyme sensors to pass the optionally pretreated with assay reagents samples through one of the measuring cells and to determine the process parameters in the flow of the sample through the measuring cell. If a solid phase-bound affinity assay is carried out in one of the measuring cells, in which analyte or other target molecules bound to immobilized receptors are provided with an enzyme label, in a flow-through method a substrate which is reacted with the enzyme label to produce a product with altered optical properties can be added to the Analyte or target molecules are transported out and the product formed weggetrportiert. The detection of the process parameter based on the optical property of the product formed from the substrate can be carried out directly in the detection area comprising the immobilized receptors or downstream of the detection area.

For determining the first and / or second parameter, in particular for determining a metabolite content, the method of standard addition can be used.

The device according to the invention or the method according to the invention makes it possible to determine CQAs, in particular product quality and / or product quantity, of a biotechnological process and, if appropriate, simultaneous determination of CPPs, e.g. the content, in particular the concentration, of one or more metabolites occurring in the process, wherein in addition parasitic effects can be automatically compensated. The method and the device can be used both in the laboratory and in the process.

The device and the method allow, in particular, the determination of a content, in particular a concentration, of a metabolite and / or of a product of a bioprocess in a process fluid sample taken from the bioprocess. The determination of the content of the metabolite or the determination of the content of the product in the liquid sample is also referred to below as the determination of the metabolite or determination of the product.

The invention described here has a number of advantages: The invention permits an automatic measurement procedure within a platform, in particular the integration and combination of metabolite analysis in an immunoanalyzer designed as a cabinet device for modularly adaptable target protein / product determination. A suitable platform is for example in DE 201010064391 A1 . DE 102010064392 A1 . WO 2012055606 A1 and WO 2012055607 A1 described in the disclosure of which reference is made in its entirety.

With the invention, a quasi-simultaneous and quasi-real-time determination of the most important, the quality characteristics affecting critical process parameters (CPPs) and at the same time the product quality relevant parameters (CQAs) possible.

The advantages of well-known laboratory methods are combined in an automated analyzer suitable for process analysis.

A direct process control and optimization by means of the control and evaluation device of the device according to the invention is possible. Alternatively, the control and evaluation device can communicate with a control device of the process.

The automation can be used to effectively eliminate inaccuracies in metabolite determination by the influence of disruptive effects, in particular by matrix effects of the sample to be analyzed (pH, viscosity, ionic strength, humic substances, colored and turbid solutions, etc.). The device is advantageously designed to: to apply the standard addition method to determine the content of a metabolite or a product. This leads to a higher measurement accuracy in comparison to the analyzers with automated metabolite determination mentioned at the beginning in Table I. When using the standard addition method, the metabolite determination of each sample is performed with its own calibration function (see below 2 ).

The invention will be explained in more detail below with reference to the exemplary embodiments illustrated in the figures. Show it:

1 a schematic representation of a device according to the invention;

2 a schematic representation for explaining the standard addition method.

In 1 a device for analyzing a sample liquid is shown. The entire sequence of an analysis measuring cycle is automated by the central control and evaluation device 3 , From a process container 1 For example, sample fluid from a process used to produce a product is passed through a suitable process port 2 removed and via liquid lines to the measuring cells ready for measurement 10 respectively. 11 transported. The process connection 2 includes a device for automatic sampling from the process container 1 which includes, for example, a sample lock and a liquid line connected to the sample lock. By means of a (in the 1 not shown) conveyor and transport device, which includes, for example, a pump) can, via the liquid line a predetermined amount of sample the measuring cells 10 and 11 be supplied. The central control and evaluation device 3 is designed to take the sample via the process connection 2 to control, for example, by controlling the sample lock and / or the conveying and transporting device.

The measuring readiness was previously determined by suitable, completely automated process steps, which include the successive passage of the assay reagents from the storage containers 7 to 9 include, guaranteed. These method steps are in the example shown here by the central control and evaluation device 3 controlled. For transporting the assay reagents and any other liquids, such as cleaning solutions and standard solutions into the measuring cells 10 . 11 The device comprises one or more liquid transport devices, such as pumps or pneumatic pressure sensors, which are provided by the control and evaluation device 3 are controllable. The optical or electrical, in each case generated by the measuring cells in the measurement of the sample optical or electrical, in correlation to the analyte concentration of the sample, the measuring signal is from the central control and evaluation device 3 detected.

The first measuring cell 10 is used to metabolite determination, thus a CPPs. It may, for example, comprise the automated operation of an enzymatic assay in solution with photometric detection (eg use of a commercial glucose assay kit, eg from Sigma Aldrich, based on glucose oxidase enzyme system or hexokinase / glucose-6-phosphate dehydrogenase enzyme system). In this case, the metabolite is thus glucose. For photometric detection, the first measuring cell 10 comprise an optical sensor, which converts the optical, photometric signal into an electrical signal, which serves as a measuring signal of the measuring cell 10 to the control and evaluation device 3 is issued. Alternatively or additionally, the second measuring cell 10 amperometric enzyme sensors (examples see Table I), which are designed to generate an electrical measurement signal and to the control and evaluation device 3 issue.

The second measuring cell 11 serves to determine a product content of the liquid sample, thus a CQA. It is preferably based on a solid-phase affinity immunosensor, for example with the in DE 201010064391 A1 . DE 102010064392 A1 . WO 2012055606 A1 and WO 2012055607 A1 described properties. In particular, the second measuring cell 11 to do so by passing one or more of the assay reagents from one of the reservoirs 7 to 9 to build a sensor matrix having a plurality of receptors to which a in the in the measuring cell 11 to be determined to be determined analyte or another target molecule to be determined, is bound selectively and specifically. The measuring cell 11 is advantageously also designed to be automatically chemically and / or electrochemically regenerated after completion of the assay, so that the sensor matrix can be automatically regenerated repeatedly in order for new measurements on newly from the container 1 be taken samples available.

The detection of the measured variable to be determined by the assay (for example, the analyte content of the sample) is detected optically, for example. For this purpose, the second measuring cell 11 comprise an optical sensor which detects a fluorescence correlated with the measured variable or an absorption correlated with the measured variable and converts it into an electrical signal which serves as the measuring signal of the second measuring cell 11 to the control and evaluation device 3 is issued.

The measuring cells may include on-site measuring electronics, which is used for a first processing, in particular for amplifying and / or digitizing the measuring signals. A further processing of the measuring signals, in particular an evaluation for determining the measured variable or variables derived therefrom, is preferred by the control and evaluation device 3 executed.

Advantageously, the first measuring cell 10 and the second measuring cell 11 operated in the flow. The measuring cells 10 and 11 For example, both the implementation of heterogeneous and homogeneous biological assays, in particular with optical detection of the measured variable serve.

The provisions with the measuring cells 10 and 11 can be done quasi-simultaneously. Additional measurements to determine the metabolites can also be made at shorter intervals than the product determination. To compensate for the measurement inaccuracies in the metabolite determination by means of the first measuring cell 10 , which result from the matrix influences of the sample to be analyzed, the method of standard addition is used in the example shown here. These include the storage containers 5 and 6 Standard solutions with two known concentrations of the analyte. In the example shown here, the control and evaluation device is used 3 automatic dosing and transport of given volumes of standard solutions to the sample to be analyzed.

These are added to the sample and in addition to the sample without standard addition using the first measuring cell 10 analyzed automatically. All measurements are made with one and the same sample matrix. The metabolite determination of each sample is carried out with its own calibration function. Changes in the slope and the zero point of the calibration line compared to the external calibration with standard solutions with different properties relative to the sample do not affect the determination. It can be a measurement with higher accuracy. All liquids get into the waste container after passing through 12 ,

Based on 2 Let's briefly explain the standard addition procedure. In addition to the measurement signal of the unknown sample without standard addition C _S , at least one further sample with added standard must be analyzed (at least S ₁ , to S _n ). The measured signals thus obtained give the respective calibration line when plotted over the added standard concentration. After extrapolation of the function, the concentration of the unknown c _sample results from the amount of the zero of the function. Prerequisite for the application of the method is that the relationship in the work area is linear.

The control and evaluation device 3 comprises a data processing device and software executable by the latter, carrying out the above methods of standard addition and analysis of liquid samples, in particular the control of the device and the determination of values of the monitored parameters from those of the measuring cells 10 . 11 supplied measuring signals, serves. Next, the control and evaluation device 3 Display means for displaying measured values and parameters of the device and input means for inputting commands or parameters by a user.

It may be configured to control a monitored bioprocess. Alternatively, it can also be configured to output the determined values of the process parameters to a controller of the monitored process. In both cases, the two are using the measuring cells 10 . 11 certain parameter values are used for different functions, namely for regulation (metabolite) and for timely monitoring of product quality.

LIST OF REFERENCE NUMBERS

1

 Process vessel with sample liquid

2

 Process connection / sampling system

3

 central control and evaluation device

4

 enclosure

5

 Stock Standard Solution 1

6

 Stock Standard Solution 2

7

 Storage 1. Assay reagent

8th

 Stock 2. Assay reagent

9

 Stock n. Assay reagent

10

 first measuring cell

11

 second measuring cell

12

 waste container

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• EP 1698891 A1 [0008]
• DE 3406223 A1 [0009]
• US 3655958 [0009]
• US 2007/0224702 A1 [0010]
• DE 19612766 A1 [0011]
• DE 102010064391 A1 [0012]
• US 2005/0208473 A1 [0013]
• US 2007/0292958 A1 [0014]
• US 2008/0241966 A1 [0015]
• DE 201010064391 A1 [0026, 0043, 0054]
• DE 102010064392 A1 [0026, 0043, 0054]
• WO 2012055606 A1 [0026, 0043, 0054]
• WO 2012055607 A1 [0026, 0043, 0054]

Cited non-patent literature

• Rodrigues, ME, Costa, AR, Henriques, M. Azeredo, J., Oliveira, R. (2010). Technological progress in monoclonal antibody production systems. Biotechnol Prog, 26 (2), p.332-351 [0003]
• Lactate, ammonium, glutamate, in general the metabolites, can be reliably determined (Rodrigues et al., Page 343, left, 4th paragraph) [0003]
• Rodrigues et al. P. 343 right 4th paragraph [0003]

Claims (15)

Device for the automated determination of at least two different process parameters of a process fluid of a process, in particular a bioprocess, comprising: A first measuring cell, which is designed to provide a first measuring signal dependent on a first process parameter of a first sample of the process liquid; A second measuring cell, which is designed to provide a second measuring signal dependent on a second process parameter of a second sample of the process fluid; and A control and evaluation device which serves to monitor and / or control the process, and which is configured to receive and process the first and the second measurement signal, in particular to determine a measured value of the first process parameter based on the first measurement signal and based on the second measurement signal to determine a measured value of the second process parameter; wherein the first measurement signal and the second measurement signal serve different functions in the context of monitoring and / or controlling the process. The apparatus of claim 1, wherein the first process parameter is a critical process parameter (CPP), and wherein the second process parameter is a product quality critical parameter (CQA) of the process. Apparatus according to claim 1 or 2, wherein the apparatus is adapted to measure the first and / or second process parameters in the flow. Device according to one of claims 1 to 3, wherein the first process parameter is a metabolite content of the sample, in particular anabolite or catabolite content, or a content of viablen and / or dead cells. Apparatus according to any one of claims 1 to 4, wherein the second process parameter is a content of the product of the process in the liquid sample or a product activity. Device according to one of claims 1 to 5, wherein the device is adapted to determine the first and / or the second parameter, in particular a metabolite content, by means of a standard addition method. Device according to one of claims 1 to 6, wherein the first or the second measuring cell for carrying out a solid-phase, immunological assay for generating their measurement signal is configured. Device according to one of claim 7, wherein the respective other measuring cell generates its measuring signal without the implementation of a solid phase-bound immunological assay, in particular using an optical or an amperometric sensor and / or a photometric cuvette test. Device according to one of claims 7 or 8, wherein the first measuring cell has at least one enzyme sensor. Device according to one of claims 1 to 9, further comprising a process connection, which in particular a device for automatic sampling from a process vessel in which the process liquid is contained and / or comprises a device for sample treatment. Apparatus according to claim 10, wherein the device for sampling is connectable to both laboratory bioreactors and process bioreactors. Method for the automated determination of at least two process parameters of a process fluid of a process, in particular of a bioprocess, by means of a device according to claims 1 to 11, wherein the determination of the first and the second process parameters are independent of each other. The method of claim 12, wherein as a first parameter, a control parameter, in particular a metabolite content, by means of an enzyme sensor and as a second parameter, a product quality-relevant parameter, in particular a product content is determined by means of an affinity assay. Method according to one of claims 12 to 13, wherein the first and the second parameter by repeatedly executable step sequences, the passage of one or more liquids, in particular of samples of the process liquid, by which measuring cells comprise, are determined several times, in particular automatically, during a course of the process. Method according to one of claims 12 to 14, wherein for the determination of the second parameter, in particular a Metabolit content, a standard addition method is applied.

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